In Vitro & In Vivo Studies on Identifying and Designing Temporin-1CEh from the Skin Secretion of Rana chensinensis as the Optimised Antibacterial Prototype Drug

Abstract

Amphibian skin secretion is an ideal source of antimicrobial peptides that are difficult to induce drug resistance to due to their membrane-targeting mechanism as a new treatment scheme. In this study, a natural antimicrobial peptide Temporin-1CEh was identified by molecular cloning and mass spectrometry from the skin secretions of the Chinese forest frog (Rana chensinensis). Through the study of the structure and biological activity, it was found that Temporin-1CEh was a helical peptide from the Temporin family, and possessed good anti-Gram-positive bacteria activity through the mechanism of membrane destruction. Seven analogues were further designed to obtain broad-spectrum antimicrobial activity and higher stability in different physiological conditions. The results showed that T1CEh-KKPWW showed potent antibacterial activity with significantly increasing the activity against Gram-negative bacteria in vitro and in vivo with low haemolysis. In addition, T1CEh-KKPWW2 showed high sensitivity to the pH, serum or salts conditions, which applied a branched structure to allow the active units of the peptide to accumulate. Even though the haemolytic activity was increased, the stable antibacterial activity made this novel analogue meet the conditions to become a potential candidate in future antimicrobial and antibiofilm applications.

Temporins: Multifunctional Peptides from Frog Skin

“…Very recently, a new Temporin 1CE family member was identified and characterized: Temporin 1CEh [ 24 ]. The peptide shows inhibitory activity, mainly against Gram-positive bacteria, but it is endowed with an elevated haemolytic activity (HC 50 = 152.6 μM).…”

“…Finally, an analogue of temporin 1CEh named T1CEh-KKPWW (KKWVPWKKIANIL-NH 2 ) was reported. It presents improved antibacterial activity against Gram-negative bacteria and low haemolysis [ 24 ].…”

Proteins extracted from pearl oyster (Pinctada martensii) with efficient accelerated wound healing in vitro through promoting cell proliferation, migration, and collagen formation

“…For instance, catfish mucus exhibits potent antimicrobial properties [ 17 ], and researchers have developed antimicrobial peptide products derived from recombinant peptides found in catfish mucus [ 18 ]. The mucus of the amphibian Rana chensinensis also demonstrates multifunctional activity, protecting the frog's survival and promoting skin healing [ [19] , [20] , [21] ]. Snail mucus is another example that has been used in treating skin wounds and promoting skin whitening.…”